BME100 s2014:W Group7 L4

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BME 100 Spring 2014 Home
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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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OUR TEAM

Name: Velia Butruz
Role(s)
Name: Matthew Welz
Role(s)
Name: Elizabeth Hansen
Role(s)
Name: Valerie McDonald
Role(s)
Name: Haley Sivertson
Role(s)
Name: student
Role(s)

LAB 1 WRITE-UP

Initial Machine Testing

The Original Design

This machine is called an OpenPCR and is used to replicate DNA. The machine goes through a continuous process of heating and cooling during the replication process. It contains a display screen that shows temperature of plate and lid. It also gives an estimated time of completion, the cycle it is on and the experiment it is. The DNA is placed into separate test tubes in the heating plate of the machine along with the primers and all of the other essentials for replication. It has a power cord and a USB cord that is used to connect to software on the computer that controls the experiment.


Experimenting With the Connections

When we unplugged the display board from the sensor/processing board, the machine display board turned off. When we unplugged the white wire that connects the processing board to the heating plate, the machine's display board showed that an inaccurate temperature reading of -44℃ after unplugging the heat sensor.


Test Run

The original test was ran on March 19, 2014. The machine passed the test run. After an hour and fifteen minutes and having gone through 21 of the 35 cycles, it was determined that the machine was working properly.




Protocols

Thermal Cycler Program


DNA Sample Set-up


DNA Sample Set-up Procedure

  1. Denaturing - Separating the two DNA strands at temperature around 95°C
  2. Annealing - Two primers bind with its complementary strand; temperature may vary based on the size of the primer.
  3. Primary Extension - The addition of new base pairs to the complementing pair already available.


PCR Reaction Mix

  • What is in the PCR reaction mix?

In the PCR reaction mix, there is Taq DNA polymerase, MgCl2, dNTP's, forward primer and reverse primer.


DNA/ primer mix

  • What is in the DNA/ primer mix?

In the DNA/primer mix, there is a sample of the patient's DNA.




Research and Development

PCR - The Underlying Technology

(Add a write-up, essay-style, organized into paragraphs with descriptive headers, based on the Q&A's from Section three of your worksheet)

(BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)